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Drought stress affects plant growth and production. To cope with drought stress, plants induced physiological and metabolic changes, serving as a protective approach under drought-stress conditions. The response to drought can vary based on plant type (C3 vs. C4) and the intensity of the stress. Therefore, here we aimed to investigate the different responses of wheat C3- Triticum aestivum and C4- Amaranthus caudatus plants to drought stress. To this end, the growth, photosynthetic parameters, oxidative stress, total antioxidant capacity, primary metabolites (amino acids and organic and fatty acids) and secondary metabolites (polyamines) were analyzed. Drought stress reduced growth, biomass, relative water content, water potential, and photosynthesis in both plants, with more severe effects observed in wheat. Drought-induced reduction in photosynthesis was linked to lower stomatal conductance, reduced photosynthetic enzyme activity, and decreased Fv/Fm, indicating impaired PSII function, effects that were more pronounced in wheat than in amaranth. This was accompanied by increased oxidative damage, as indicated by elevated levels of lipid peroxidation. To cope with drought stress, both plants accumulated metabolites involved in antioxidant defense and osmoregulation, including total antioxidant capacity, soluble sugars, proline, polyamines, organic acids, and fatty acids. This response was more pronounced in wheat, indicating its active deployment of defenses to cope with significant stress, in contrast to Amaranthus ' greater physiological resilience.

The need for improving plant production, nutritional value, and medicinal applications has become increasingly important due to the growing global population. The caraway ( Carum carvi L) plant has been recognized for its broad range of nutritional and therapeutic uses. Consequently, this study aimed to increase caraway seeds’ nutritional and biological value. To achieve this, the Actinopolyspora sp. JTT-0 strain, isolated from the medicinal plant Tephrosia purpurea , was investigated for its potential biofortification role to enhance caraway yield and quality. Our results revealed significant improvements ( p  < 0.05) in various physical parameters, such as seed yield, pod length, and bulk density, in the treated seeds compared to the controls. Along with the yield increase, there were notable elevations in primary metabolites such as total sugars, proteins, and amino acids. Furthermore, secondary metabolites, including essential oils (EOs), alkaloids, steroids, phenols, and vitamins (e.g., tocopherol and ascorbic acid), also showed significant increases. Notably, the EO constituents showed varying levels of enhancements, with the highest increases in β-pinene (186.2%) and carvacrol (49.2%). Moreover, the treated seeds exhibited improved biological activity, as evidenced by their anti-oxidant (anti-lipid peroxidation and DPPH assays) and anti-microbial properties compared to the controls. The study reported a positive biofortification effect of the Actinopolyspora sp . JTT-01 strain on enhancing caraway seed’s quality and yield. However, additional field trials are needed to evaluate the commercial biofertilization capacity of this strain for caraway and other plants.

Although drought responses of C3 and C4 plants have been widely investigated, direct comparative analyses of their physiological and detailed metabolic biosynthetic adaptations remain limited. This study provided a detailed pathway-level analysis of drought stress responses in a Brassica oleracea ( kale, C3) versus Echinochloa crus-galli (barnyard grass, C4) moving beyond previous comparisons that typically focus on broad physiological differences. Our study integrates multi-level physiological, molecular, biochemical, and metabolic data, emphasizing specific drought-adaptive biosynthetic pathways such as anthocyanin and glucosinolate synthesis. Drought stress significantly reduced biomass and photosynthetic efficiency ( p  < 0.05) in both species, with kale exhibiting greater declines. Kale also accumulated higher oxidative stress indicators, with H₂O₂ and lipid peroxidation (MDA) increasing by 21.81% and 113%, respectively, relative to barnyard grass, indicating stronger oxidative damage. Both species significantly upregulated antioxidants metabolites and enzymes, reflecting activation of protective mechanisms. Anthocyanin content rose by 83% in barnyard grass and 113% in kale under drought. Detailed pathway analysis revealed elevated precursor levels (phenylalanine and p-coumaroyl-CoA) and increased activities and gene expression of key enzymes in the phenylpropanoid pathway, including cinnamate-4-hydroxylase (C4H), chalcone synthase (CHS), and 4-coumarate-CoA ligase (4CL), highlighting the specific C3-C4 photosynthetic-type biochemical strategies for drought adaptation. The strong activation of defense pathways such as anthocyanin and glucosinolate biosynthesis suggests that kale compensates for its lower drought tolerance by increasing metabolic flexibility. Molecular evidence further supports induced expression of anthocyanin related biosynthetic genes. The detailed, multi-level metabolic analysis uncover the -specific C3-C4 photosynthetic-type biochemical strategies for drought adaptation. This work advances plant physiology knowledge by revealing how C3 and C4 plants differ in metabolic drought adaptations, offering practical relevance for both crop improvement and weed management. This reveals key physiological and biochemical differences, with kale (C3) needing more metabolic adjustment to handle drought.

Basil ( Ocimum L.) is an important essential oil crop, medicinal plant, and culinary herb, belonging to the Lamiaceae family. It has extensive nutritional and therapeutic benefits, making it valuable in culinary and medicinal applications. Hence, in this study, we aimed to induce basil’s nutritive and biological value. To this end, this study evaluates the potential of the plant growth-promoting Pseudomonas JP0825, isolated from the Jazan region, KSA, as a biopriming agent to improve the growth, nutritional quality, and bioactive compound profile of sweet ( Ocimum basilicum L.) and American ( Ocimum americanum L.) basil. The molecular identification of Pseudomonas JP0825 confirmed its phylogenetic relationship with other beneficial Pseudomonas species. Our findings revealed significant increases in photosynthetic pigments, biomass, and proximate composition, particularly in sweet basil, following inoculation. Elevated levels of vitamins, amino acids, and organic and fatty acids were observed, alongside enhanced secondary metabolites like phenolics and flavonoids, correlated with enhanced antioxidant and antimicrobial activity. The antioxidant properties of treated basil improved significantly, as indicated by increased FRAP and ABTS activities. Furthermore, Pseudomonas JP0825 demonstrated an ability to boost the antimicrobial activity against various pathogenic bacteria and fungi, including Staphylococcus epidermidis , Enterococcus faecalis , Salmonella typhimurium , and Aspergillus flavus . These findings highlight the strain’s potential as a sustainable alternative to chemical inputs, offering improvements in crop quality and resilience, and contributing to global food security efforts.

This study investigated the effects of non-thermal atmospheric plasma (NTAP) treatment on the growth, chemical composition, and biological activity of geranium ( Pelargonium graveolens L’Herit) leaves. NTAP was applied at a frequency of 13.56 MHz, exposure time of 15 s, discharge temperature of 25 °C, and power levels (T1 = 50, T2 = 80, and T3 = 120 W). Results demonstrated significant increases ( P  < 0.05) in fresh and dry biomass at all treatment levels compared to control, with the highest improvements seen in T3. Mineral content (K, P, Ca, Fe, Mg, Zn, and N) was significantly elevated, particularly at T3. Chlorophyll content (a + b and carotenoids) also showed marked increases across all treatments, correlating with enhanced p hotosynthetic rates. Improved photosynthesis led to enhanced accumulation of primary metabolites, such as amino acids, organic acids, and fatty acids. NTAP treatments, mainly T3, significantly increased levels of essential and non-essential amino acids, oxalic, isobutyric, and fumaric acids. They also enhanced unsaturated fatty acids, such as oleic acid (C18:1), and saturated fatty acids, including myristic (C14:0) and stearic (C18:0). These improvements provided precursors for the synthesis of secondary metabolites, particularly phenolics. The increased phenolic content in turn explained the improved antioxidant capacity observed in Fluorescence Recovery After Photobleaching FRAP, anti-lipid peroxidation, superoxide radical scavenging, and hydroxyl radical scavenging assays, especially at T2 and T3 treatments. Antimicrobial activity was elevated across all treatments, with the T3 treatment notably inhibiting all tested bacterial and fungal strains, particularly Sarcina lutea. In conclusion, NTAP treatment significantly improved growth, biomass, and the phytochemical profile of geranium leaves, enhancing their antioxidant and antimicrobial properties, thereby increasing the potential nutritional and therapeutic value of the plant.

The present research aimed to assess the metabolomic responses of wheat to olive mill wastewater (OMWW) and drought stress treatments. Wheat plants were cultivated under controlled conditions with the following treatments: control (75% field capacity, FC), OMWW (75 ml L −1 ), drought stress (40% FC, applied 30 days after sowing), and a combined treatment of OMWW and drought stress. Drought stress alone reduced grain yield by 67%, while the OMWW-treated plants resulted in a 29% reduction under stress relative to the control. OMWW application improved soil properties, enhancing organic matter and nutrient levels. Wheat grains from OMWW-treated plants exhibited higher sugar content and related enzyme activities, indicating improved metabolism, with significant increases in starch, fructose, and glucose levels alongside stable invertase and sucrose phosphate synthase activities. The study also noted substantial changes in amino acids, fatty acids, and phenolic acids in plants subjected to OMWW and drought stress. These modifications indicate OMWW’s capability to influence vital biochemical pathways and boost antioxidant capacities in wheat. In conclusion, OMWW proves to be an effective soil amendment that mitigates drought stress and contributes to the production of nutrient-rich, resilient wheat, underscoring its potential as a sustainable agricultural practice in water-scarce areas.

Anise (Pimpinella anisum L.) seeds have various nutritional and therapeutic benefits and are thus considered a valuable addition to animal and human health. Hence, in this study, we aimed to induce the nutritive and biological value of anise seeds. To this end, the potential biofortification effect of the endophytic Actinomycetota sp. JW0824 strain, isolated during the fall of 2023 from the medicinal plant Achyranthes aspera, exhibiting natural distribution in the Jazan region of Saudi Arabia, was investigated in four varieties of anise seeds from Egypt, Tunisia, Syria, and Morocco. Results revealed significant increments (p < 0.05) in the seed dry weight percentage (DW%) and oil yields. In line with increased biomass accumulation, the metabolism of the primary and secondary metabolites was increased. There were differential increases in proteins, sugars, flavonoids, alkaloids, phenols, vitamins (e.g., β-carotene, ascorbic acid), and essential oil components (e.g., phenylpropanoids and monoterpenes), along with their precursor phenylalanine. Consistently, the activity of L-phenylalanine aminolyase (PAL) was increased in the Egyptian and Tunisian varieties at 83.88% and 77.19%, respectively, while 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase (DAHPS) activity increased in all varieties, with a significant 179.31% rise in the Egyptian variety. These findings highlight the beneficial effects of Actinomycetota sp. JW0824 as a bioinoculant for anise seeds, suggesting its potential application in agricultural practices to improve seed yield and quality. Further field trials are recommended to assess the commercial viability of this endophyte for enhancing anise seed production and potentially benefiting other plant species.

Background: The genus Ephedra (Ephedraceae) is a dioecious gymnosperm, where female individuals produce the pharmacologically active ephedrine alkaloids. Identifying the sex of specimens without reproductive cones is challenging due to their xeromorphic and morphological similarity. The challenges in sex identification complicate conservation and propagation efforts. Methods: Sequence-Related Amplified Polymorphism (SRAP) markers were applied to distinguish genders in five Ephedra species, particularly the vegetative branches, as well as powdered and fragmented specimens. The fresh material for the five studied Ephedra species and two sexes per species (totaling 10 samples; 5 females & 5 males) was collected from Sinai, Egypt. Results: The SRAP marker results revealed an exclusively male-specific band, and this is not applicable in females in the studied species. The applied SRAP markers grouped males and females in different UPGMA clusters and proved their efficiency in distinguishing between males and females in the five studied species. The Polymorphic Information Content (PIC) values are low (0.16–0.38); this suggests moderate genetic diversity between the females of the studied species, reflecting slow evolutionary rates. Conclusions: The SRAP markers are efficient for identifying Ephedra species at the species and gender levels, even in the absence of sex organs and molecular sequences. Recommendation: This study recommends the use of SRAP markers for conserving and propagating female plants for ephedrine production and suggests sequencing a 95 bp male-specific band to determine if it corresponds to a known sex-linked gene.

The genus Tourn. ex L. (Ephedraceae) is an important source with pharmacological and environmental potential. Conversely, spp. still exhibit taxonomic complexity, especially for the specimens lacking reproductive cones. This complexity is attributed to its xeromorphic features, notably the reduced leaves and analogous assimilating branches, which make the species identification a real challenge. The current study provides a pioneering approach to distinguish fragments of species at the gender level. This study was based on the stem anatomy and stem epidermal features using a light microscope for five species ( Decne., Forssk., Fisch. & C.A.Mey., Forssk. and Boiss.) represented by ten genders collected from S. Sinai, Egypt. Anomocytic and brachyparacytic stomata, tanniniferous idioblasts, annual rings, a terete and furrowed outline, the number and width of tracheids, patches of cortical fibers, unicellular trichomes, druses, solitary crystals, and the activity of interfascicular cambium were among the distinguishing features that were found. Different statistical analyses were applied to explore the diversity at interspecific and intra-generic levels. This study revealed that the stem anatomy was not only an efficient tool for identifying the investigated five species at the species level but also presented a differential key to distinguish between genders and species. In addition, our results indicated that the epidermal features played a critical role in differentiating the studied species at the gender level. This study confirms the efficacy of stem anatomy as an identification approach for the species at the gender level and recommends this approach to identify the fragmented for taxonomical, pharmaceutical, and medical applications.

The genus Ephedra Tourn. ex L. (Ephedraceae) still exhibits taxonomic complexity that has not yet been resolved. This study aimed to determine the taxonomic identity of the Ephedra species in Egypt and identify the fatty acid profile and its diversity at the gender level as a taxonomic tool for specimens lacking reproductive cones. The current study provides a pioneering approach that distinguishes Ephedra species at the gender level. A total of 120 fresh individuals were collected from 20 locations representing different habitats where Ephedra plants grow in Egypt. In addition, herbarium specimens were deposited in Egyptian herbaria. The studied morphological traits included 30 vegetative characteristics and 72 traits of the reproductive organs of both genders. The fatty acid content was measured using gas–mass chromatography (GC-Mass). The taxonomic revision revealed that the Genus Ephedra was represented in the Egyptian flora by five species, Ephedra alata in section Alatae and E. aphylla, E. ciliata, E. foemina, and E. pachyclada in section Ephedra. South Sinai hosts these five species and represents the center of diversity for this genus in Egypt. The vegetative characteristics were subjected to principal component analysis (PCA), which revealed a distinct separation of the five studied species. Similarly, the cone traits treated by hierarchical clustering revealed intra-gender variations. The taxonomic key was developed based on the morphological traits to distinguish the studied species at the gender level. In total, 51 fatty acids were identified from the studied species and grouped as 18 saturated, 16 monounsaturated, and 17 polyunsaturated fatty acids. In the absence of reproductive cones, the lipid content and fatty acid composition of the vegetative parts displayed significant interspecific and intra-gender variations. Therefore, fatty acids can be used to efficiently identify the studied species when they lack reproductive cones. This study proved the efficacy of a multidisciplinary approach to identify Ephedra species at the gender level and recommends this trend for future studies of this genus.